RGD-conjugated In Situ forming hydrogels as cell-adhesive injectable scaffolds

Abstract

In this study, a bioactive hydrogel was prepared from a chitosan derivative and Arg-Gly-Asp (RGD)-conjugated polypseudorotaxane, which is a cell-adhesive extracellular matrix. Chitosan was modified with 4-hydroxy phenyl acetic acid to obtain a water-soluble product for enzymatic cross-linking. Tyramine-terminated polypseudorotaxane (PRx) was prepared from the inclusion complex of a tyramine-terminated poly(ethylene glycol) backbone, and α-cyclodextrin (α-CD). Gly-Arg-Gly-Asp-Ser (GRGDS) was conjugated to the PRx using 4-nitrophenyl chloroformate (NPC) and partially carboxylated with succinic anhydride. The structure of the PRx-RGD and 4-hydroxylphenylacetamide chitosan (CHPA) was characterized by 1H NMR and FTIR spectroscopy. The RGD content in PRx-RGD was determined to be 0.19%. PRx-RGD and CHPA solution was crosslinked to form a bioactive hydrogel in the presence of horseradish peroxidase (HRP) and hydrogen peroxide (H2O2), which exhibited rapid gelation (∼20 sec). An in vitro cell culture was carried out with L929 mouse fibroblasts for 1 and 3 days. The results showed that fibroblasts adhered better and appeared to be more biocompatible on the RGD-conjugated hydrogel than the hydrogel without RGD. The combined results highlight the potential use of this bioactive hydrogel as an injectable scaffold in tissue engineering applications. Open image in new window